Mass Spectrometry-Based Proteogenomic Approaches Reveal Insight into the Activities and Functions of Microbial Isolates and Communities*
Robert Hettich, Ritin Sharma, Xiaoxin Liu, Karuna Chourey, Chongle Pan, and Richard Giannone
Oak Ridge National Laboratory, Oak Ridge, TN, U.S.A.
With the availability of complete microbial genome sequences for cultured organisms as well as more complex environmental samples, systems biology in microbial communities is becoming feasible by combining genomic, transcriptomic, proteomic and metabolic information. Our current work seeks to develop and demonstrate advanced “shotgun” mass spectrometry techniques for the comprehensive characterization of microbial proteomes. The goal of this research is to enable a detailed glimpse into the functional state and metabolic activities of microbial systems. The approach is based on multidimensional liquid chromatography interfaced on-line with tandem mass spectrometry. A variety of mass analyzers are employed, ranging from quadrupole ion traps (high throughput) to Orbitraps and Fourier transform ion cyclotron resonance instruments (high performance, accurate mass measurements). We have focused on integrated experimental/bioinformatic approaches for both qualitative and quantitative proteome determinations. We have developed a detergent based proteome extraction method which can be combined with high performance LTQ-Orbitrap-Velos measurements to investigate natural microbial communities, such as those involved in carbon cycling in terrestrial (soils, sediments) as well as aqueous (acid mine drainage, deep sea methane seeps) systems. We have extended these MS-based techniques to characterize a variety of microbial species that are involved in bioremediation, bioenergy, human microbiome health, and natural environmental communities.
This seminar is hosted by Dr. Mike Henson.
As usual, all seminars in our series are free and open to any interested public. Please visit our seminar calendar for more information and other events. http://www.clemson.edu/cafls/departments/biosci/research/seminars.html
Dr. Charlie Wei
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